FCA UNICAMP LABORATÓRIO DE LÁCTEOS PROBIÓTICOS PREBIÓTICOS NOVEMBRO DE 2020 FACULDADE DE ENGENHARIA DE ALIMENTOS UNICAMP - CAMPINAS INTERNATIONAL WORKSHOP ON ADVANCES OF PROBIOTICS AND PREBIOTICS (IWAPP) PRE E PROBIÓTICOS PARA SAÚDE E EQUILÍBRIO DE HUMANOS, ANIMAIS E AMBIENTE teórico workshop prático livro de resumos
152
Embed
livro de resumoslivro de resumos NOVEMBRO DE 2020 FACULDADE DE ENGENHARIA DE ALIMENTOS | UNICAMP - CAMPINAS FCA UNICAMP LABORATÓRIO DE LÁCTEOS …
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
FCAUNICAMP
LABORATÓRIODE LÁCTEOSPROBIÓTICOSPREBIÓTICOS
NOVEMBRO DE 2020 FACULDADE DE ENGENHARIA DE ALIMENTOS
UNICAMP - CAMPINAS
INTERNATIONAL WORKSHOPON ADVANCES OF PROBIOTICS
AND PREBIOTICS (IWAPP) PRE E PROBIÓTICOS PARA SAÚDE E EQUILÍBRIO DE
HUMANOS, ANIMAIS E AMBIENTE
t e ó r i c oworkshopp r á t i c o
livro de resumos
NOVEMBRO DE 2020FACULDADE DE ENGENHARIA DE ALIMENTOS | UNICAMP - CAMPINAS
FCAUNICAMP
LABORATÓRIODE LÁCTEOSPROBIÓTICOSPREBIÓTICOS
INTERNATIONAL WORKSHOPON ADVANCES OF PROBIOTICS
AND PREBIOTICS (IWAPP)
INTERNATIONAL WORKSHOPON ADVANCES OF PROBIOTICS
AND PREBIOTICS (IWAPP)
ORGANIZAÇÃO
FCAUNICAMP
LABORATÓRIODE LÁCTEOSPROBIÓTICOSPREBIÓTICOS
APOIO
The DuPont Logo is a trademarks ofDuPont de Nemours, Inc., or its af�liates.
Fundo de Apoio ao Ensino, à Pesquisa e à Extensão
AEPEXF
INTERNATIONAL WORKSHOPON ADVANCES OF PROBIOTICS
AND PREBIOTICS (IWAPP) PRE E PROBIÓTICOS PARA SAÚDE E EQUILÍBRIO DE
HUMANOS, ANIMAIS E AMBIENTE
t e ó r i c oworkshopp r á t i c o
resumos detrabalhos científicos
FCAUNICAMP
LABORATÓRIODE LÁCTEOSPROBIÓTICOSPREBIÓTICOS
NOVEMBRO DE 2020FACULDADE DE ENGENHARIA DE ALIMENTOS | UNICAMP - CAMPINAS
ORGANIZAÇÃO
FCAUNICAMP
LABORATÓRIODE LÁCTEOSPROBIÓTICOSPREBIÓTICOS
APOIO
The DuPont Logo is a trademarks ofDuPont de Nemours, Inc., or its af�liates.
Fundo de Apoio ao Ensino, à Pesquisa e à Extensão
AEPEXF
NOVEMBRO DE 2020FACULDADE DE ENGENHARIA DE ALIMENTOS | UNICAMP - CAMPINAS
FCAUNICAMP
LABORATÓRIODE LÁCTEOSPROBIÓTICOSPREBIÓTICOS
INTERNATIONAL WORKSHOP ON ADVANCESOF PROBIOTICS AND PREBIOTICS (IWAPP)
24/11 Terça-feira/ Tuesday08:30 - AberturaManhã – Plenárias (food / vet / ambiental) / AM - Plenary ( food / vet / environmental)08:35h - 09:05h (+5min)Palestrante internacional Dr. Arthur Ouwehand (Dupont/Finlândia)."Paraprobiotics and postbiotics for food applications"09:10h - 09:40h (+5min)Palestrante internacional Profa. Dra. Emma Allen-Vercoe (University of Guelph/Canadá). “Human gut-derived microbes and ecosystems as novel probiotics and drugs ”09:45h - 10:00h Intervalo10:00h - 10:30h (+5min)Palestrante internacional Prof. Dr. Ben Willing (University of Alberta/Canadá)."Probiotics instead of antibiotics for animal productions and bene�ts to animals and humans"
09:05h - 09:35 (+5min)Palestra 2Profa. Dra. Elane Prudêncio (UFSC)“Leites de outras espécie como matrizes para probióticos e prebióticos”
09:40h - 10:10h (+5min)Palestra 3Prof. Dr. Erick Esmerino (UFRJ)“Métodos Sensoriais para desenvolvimentos de Alimentos e Probióticos” 10:15h - 10:45hIntervalo e visita virtual de resumos cientí�cos10:45h - 11:15h (+5min)“Benefícios dos Prebióticos isolados e combinados para uma maior saúde animal com e sem desa�o com Salmonella entericaDra. Juliana Bueno - Empresa Yes11:20h - 11:50h (+5min)Palestra 4Profa. Dra. Glaucia Pastore (UNICAMP)“Prebióticos – nova de�nição e abrangência”
Vet08:30h - 09:00h (+5min)Palestra 1:Prof. Dr. Raphael Lucio Andreatti Filho(UNESP), Campus Botucatu. “Probióticos e controle de Salmonella em avicultura” 09:05h - 09:35 (+5min)Palestra 2Prof. Dr. Marcio Antonio Brunetto. USP“Pre e probióticos para PET”
09:40h - 10:10h (+5min)Palestra 3Prof. Cesar Augusto Pospissil Garbossa (USP)"Uso de prebióticos e probióticos em substituição aos antibióticos e manipulação do microbioma em suínos"10:15h - 10:45hIntervalo e visita virtual de resumos cientí�cos10:45h - 11:15h (+5min)“Uso de probióticos em animais de produção” - Vinícius TeixeiraEmpresa Evonik
11:20h - 11:50h (+5min)Palestra 4Prof. Dr. Ricardo Pinheiro de Souza Oliveira (USP)“Bacteriocinas sintetizadas por culturas probióticas e suas aplicações em animais de produção”
Amb / Env 08:30h - 09:00h (+5min)Palestra 1Prof. Dr. Everlon Cid Rigobelo (UNESP)“Uso de Lactobacillus acidophilus como promotores de crescimento de plantas”
09:05h - 09:35 (+5min)Palestra 2Profa. Dra. Fabiana Pilarski (UNESP)“Impacto ambiental da utilização de bactérias probióticas na água de piscicultura.”09:40h - 10:10h (+5min)Palestra 3Profa. Dra. Maria Rita Muzzi (UFMG)“Uso de bactérias probióticas para recuperação de áreas degradadas”
10:15h - 10:45hIntervalo e visita virtual de resumos cientí�cos10:45h - 11:15h (+5min)Palestra empresa convidada
11:20h - 11:50h (+5min)Palestra 4Dr. Rodrigo Mendes (EMBRAPA)“Bactérias probióticas com membros da microbiota do solo”
25/11 Quarta-feira / Wednesday
Área Food Área Vet
11:45h Encerramento
AGENDA
10:35h - 12:00h Microencapsulação de probióticosProfa. Dra. Ana Silvia Prata Soares (UNICAMP), Profa. Dra. Carmen Fávaro Trindade (USP), Ms. Augusto Tasch Holkem, Ms. Marluci Palazzolli da Silva, Ms Edison Motta dos Santos Junior, Ms Fernanda Sviech
10:35h - 12:00h Obtenção de ração para peixes e suínos contendo probióticosProfa. Dra. Maria Teresa Clerice(UNICAMP), Dr. Leonardo Tachibana (IP/APTA), Dr. Fábio Enrique Lemos Budiño (IZ/APTA),Dra. Danielle de Carla Dias (IP/APTA), Ms Ulliana Marques Sampaio
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
5
ACTIVIDAD ANTIMICROBIANA DE LEVADURAS DE ORIGEN VITIVINÍCOLA
SELECCIONADAS CON APTITUDES PROBIÓTICAS.
Vergara Silvia1,2, Leiva M. Jose1,2, Mestre M. Victoria1,2,3, Nally M. Cristina1,2,3, Kuchen
Artichoke (Cynara cardunculus L.), a plant native to the Mediterranean basin and
belonging to the Asteraceae family, has been used since ancient times as a natural food and
herbal medicine, but its leaves are usually discarded. Notably, these leaves are rich in inulin, a
natural prebiotic. Prebiotics are non-digestible but fermentable foods that beneficially affect the
host by selectively stimulating the proliferation or activity of desirable bacteria populations in
the colon. Inulin has bifidogenic, hepatoprotective and antidiabetic effects and stimulates the
absorption of minerals such as calcium and magnesium, among others benefits. The tinctures
of artichoke leaves (ALT) rich in inulin were obtained by extraction with pressurized liquid
(PLE) using intermittent solvent purging. Ethanol (99.5%) was used as solvent in the PLE, and
four control variables were studied, so a 24-1 factorial design was adopted to evaluate the effect
of each process variable on the content of inulin in the extracts. The studied parameters were
temperatures of 40, 50 and 60 °C, solvent volumes of 60, 70 and 80% of the extraction cell
volume, contact times between the solvent and the matrix in each cycle of 6, 8 and 10 min
(solvent purging time) and number of cycles 3, 4 and 5. Thin layer chromatography (TLC) was
performed to check the presence of inulin using aluminum chromatography sheets TLC silica
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
22
gel 60 F254 as a stationary phase. A Propanol-Butanol-Water solution (12:4:3) was used as the
mobile phase and the tinctures were diluted in distilled water at a concentration of 200 mg/mL.
Inulin content (IC) was quantified by spectrophotometry. TLC showed inulin presence in all
ALT. The highest IC (109.53 mg inulin/g) was obtained when the extraction was conducted at
60 °C, with 60 % of the extractor volume, 6 min of purging time for 5 cycles. The lowest IC
(38.29 mg inulin/g was measured in the ALT extracted at 40 °C, with 60 % of the extractor
volume, 6 min and 3 cycles. Statistical analysis via ANOVA and Pareto diagram showed that
none of the independent variables had a significant effect on the IC for the range of variable
values studied. The conclusion is therefore that the ALT obtained by PLE with ethanol are rich
in IC even without identifying extraction variables that have a significant influence on its
content.
We would like to thank FAPESP, process nº 2016 / 13460-8 and CAPES, financing code
001.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
23
HEMATOLOGICAL STUDY OF THE EFFECTS OF PROLONGED
ADMINISTRATION OF Β-GLUCAN ON AN IMMUNITY, IN TILAPIA,
OREOCHROMIS NILOTICUS.
Camino Fierro-Castro1*, Ingrid Camargo dos Reis2, Basia Schlichting Moromizato2, Thais
Cristina de Oliveira2, Giovanni Sampaio Gonçalves3, Jaqueline Dalbello Biller2
1Departmento de Biología celular, Fisiología e Inmunología, Fac. Biociences, Universitat
Autònoma de Barcelona, Bellaterra, 08193 Spain. *E-mail:[email protected] 2Depatamento de Produção Animal. Faculdade de Ciências Agrárias e Tecnológicas, UNESP,
Campus de Dracena; Rod Cmte João Ribeiro de Barros, km 651- Dracena/SP, 17900-000,
Brasil. 3SAA/APTA/Instituto de Pesca - Centro Avançado de Pesquisa Tecnológica do Agronegócio
do Pescado Continental, São José do Rio Preto /S,P 15025-970, Brasil.
Is well known that immune-modulation of fish with β-glucans, using injection, dietary or
even immersion, has stimulating effects on both intestinal and systemic immunity. Although
the exact effects can be variable depending on fish species, administration route or time of
administration. The aim of this study was determining the best time line to administrate the β-
glucan diet studying the effects of dietary β-glucan supplementation (10g kg-1 of β-glucan), on
immune response, and antioxidant capacity of Tilapia, Oreochromis niloticus, under over time.
For this purpose, the tilapias were fed with a control diet for 90 days and an experimental
diet for 30, 60 and 90 days to verify the existence of a prolonged effect of β-glucan over time.
Fish from all treatments were sampled, anesthetized in clove oil solution (1 g 10 L-1 of water),
and blood was collected from 12 fish (n=12) from each box (4 boxes per treatment) to evaluate
hematological variables. Hemoglobin (HGB) content was assayed by cyanomethaemoglobin
determination. Wintrobe indexes such as mean corpuscular volume (MCV), corpuscular
hemoglobin means (CHM), and mean corpuscular hemoglobin concentration (MCHC) was
derived from the primary indexes. HGB concentrations and hematocrit did not differ among
treatments, but were within normal range for species, indicating that administration of β-glucan
does not alter fish energy production. Red blood cells (RBC) seems decreased after 60 and 90
days with β-glucan diet while MCV increased the same days. Given that the hemoglobin and
hematocrit values remain stable, this could be indicating the increase in the leukocyte
population involved in the immune defense system. This theory is reinforced with the study of
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
24
leukocyte respiratory activity Burst (NBT assay in blood) that show a significantly increases
after 90 days with a β-glucan diet. The knowledge of the hematological response under different
diets can be a useful tool for improve the administration of β-glucan and make it really effective.
Grants: The São Paulo Research Foundation - FAPESP – Process number:2012/22016-3.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
25
CAGAITA JUICE (EUGENIA DESYNTERICA) FERMENTED WITH KEFIR:
PHYSICOCHEMICAL AND MICROBIOLOGICAL CHARACTERISTICS
Daiane Costa dos Santos1, Tainara Leal de Sousa1, Railany Vieira Santana2, Juliana Riso
The inclusion of prebiotic from Saccharomyces cerevisiae cell wall, rich in mannan
oligosaccharides (MOS), benefits intestinal mucosa of monocavitary animals (Chacher et al.,
2017). MOS favors other species and, in addition, there is a lack of studies using ultrasound
assessment of the gastrointestinal tract (GIT) as investigating method of nutraceuticals effects.
In this way, this study evaluated whether supplementation with dry hydrolyzed Saccharomyces
cerevisiae (Sc) promotes morphometric changes in intestines of pregnant Saanen goats that are
visualized by ultrasound evaluation. Sixteen goats were separated in control (Ctr, n = 8) and
supplemented (Trat, n = 8) groups, supplementation at a dose of 7 g/animal/day, from the 30th
day of gestation until parturition. Measurements were made via ultrasound exam (USE) of the
thickness: wall of the descending duodenum (DUO), wall of the small intestine in the jejunum
or ileum (JEI) (the two organs cannot be distinguished in the USE) and wall of the large
intestine (LI) , all obtained on the right flank. From the 4th week, USE was performed on goats
(n = 16) in periodic evaluations (5, 6, 8, 10, 12, 14, 16, 18 and 20 weeks of pregnancy). Split-
plot experiment (repeated over time) was used in completely randomized balanced design with
95% confidence interval. Analysis of variance – ANOVA – was followed by Tukey test, at 5%
probability, to compare means (RStudio®, version 3.6.2.). JEI measurements were thicker in
Trat (p <0.05) at week 10 (1.18mm in Ctr; 1.57mm in Trat) and at week 16 (1.48mm Ctr;
2.00mm Trat). LI thicknesses showed different means (p<0.05) in weeks 4 (0.81mm Ctr;
1.08mm Trat) and 12 (1.08mm in Ctr; 1.69mm in Trat) and 16 (1.77mm Ctr; 1.23mm Trat). In
the fifth gestational week, the difference cannot be explained by the action of the Sc because
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
33
prebiotics can take 7 to 14 days to produce effects (Gibson et al., 2017). The detected changes
indicate that the jejunum and ileum and large intestine are segments of the GIT that are sensitive
to the action of prebiotics in adult ruminants, and that, although subtle, such differences are
verifiable by ultrasonography. These results are pioneers in ultrasound research with ruminants
and may contribute to future investigations in the morphological evaluation of in vivo GIT in
farm animals, respecting the 3Rs: Replacement, Reduction and Refinament.
Keywords: small ruminants, ultrasound, intestine, pregnancy Chacher M, Kamran Z, Ahsan U, Ahmad S, Koutoulis K, Din HQU, Cengiz Ö (2017) Use of mannan oligosaccharide in broiler diets:
an overview of underlying mechanisms. World's Poultry Science Journal 73(4):831-844.
Un microorganismo para ser considerado potencial candidato a probiótico debe
sobrevivir a las condiciones restrictivas del tracto gastrointestinal (TGI), preferentemente
adherirse temporalmente a la mucosa intestinal, y ejercer posibles beneficios para la salud del
huésped. Objetivo:evaluar propiedades relacionadas con la adhesión a células epiteliales del
TGI en 15 aislamientos de levaduras vínicas nativas. Se determinó la hidrofobicidad como la
capacidad de los microorganismos para unirse a hidrocarburos. Las levaduras activas se lavaron
3 veces con PBS 0,1M (4000rpm, 10min) y se determinó el valor DO (600nm) a cada
suspensión levuriana (concentración inicial 5x107 células/mL, T:0). Posteriormente, se añadió
el hidrocarburo xileno (1/3) durante 2min a 3500 rpm. La mezcla se incubó estáticamente
durante 1h a 37°C. Se retiró el xileno y se midió DO (600nm) de la fase acuosa. Para la
determinación de autoagregación las levaduras activadas se lavaron dos veces con PBS 0.1M.
Posteriormente, se suspendieron en PBS 0.1M (5x107 cél/mL) se agitaron 10seg e incubaron
estáticamente a 37°C. A 1mL de la fase superior se determinó DO(600nm) a 0, 2, 4 y 24h. Para
evaluar la producción de biofilm se añadió una suspensión de levaduras activas (1x106 cel/mL)
en medio YEPD. Se incubó estático por 28h a 37°C. Posteriormente, se retiró el medio, las
células fueron lavadas con PBS y dejo por 2min a temperatura ambiente; seguidamente se
agregó solución de cristal violeta (1%) por 3min, después fue retirado y las células fueron
lavada con agua destilada. El colorante adherido se eluyó con etanol (96%), y parte del mismo,
se transfirió a una placa nueva para medición de DO (620nm). Los porcentajes de
hidrofobicidad fueron variables (entre 1,05% y79,67%). Se observó que las cepas de Pichia
kudriadzevii (Pb50, Pb51, Pb52, Pb54) registraron los valores más altos (50.2-72.4) junto con
Hanseniaspora guillermondii PB15, mientras que el resto de las cepas obtuvieron porcentajes
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
43
menores al 44%. Con respecto a la autoagregación, se encontraron diferencias significativas
entre los datos de las primeras horas (2-4h) y los últimos datos (24h) (p =0,05). A las 2h, P.
kudriadzevii PB48, Pichia occidentalis PB56-PB58 y Torulaspora delbrueckii PB91
registraron valores superiores al 35%, mientras que el resto obtuvo valores inferiores al 20%,
sin cambios significativos a las 4h. Se observó un aumento entre 68% y 94% en todos los
aislamientos después de 24h de incubación. Nuestros resultados muestran que la capacidad de
autoagregación entre 2 y 4h fue variable y dependiente de la cepa. En cuanto a los valores
obtenidos a las 24h, los valores fueron más contantes. Las cepas de Hanseniaspora uvarum
PB18, T. delbrueckii PB91, Wickeranomyces anomalus PB97 podrían considerarse bajas
productoras de biopelículas teniendo en cuenta el valor umbral obtenido (A620=0.11). Candida
Sake Pb7 y P. kudriadzevii PB50 y PB53 registraron mayor formación de biofilm (>0.43). El
resto de las cepas mostraron una moderada capacidad de formación de biopelículas (>0.11-
<0.43). Los hallazgos obtenidos hasta el momento son prometedores, relevantes y direccionan
los estudios de estas cepas para futuros test que profundicen en las aptitudes y propiedades
benéficas relevantes para los microorganismos probióticos.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
44
EVALUATION OF ANTIOXIDANT RESPONSE AFTER PROLONGED
ADMINISTRATION OF Β-GLUCAN ON TILAPIA, OREOCHROMIS NILOTICUS.
Jaqueline Dalbello Biller1*, Ingrid Camargo dos Reis1, Eduardo Antônio Oliveira Cavalheiro1,
Victor Negri Libório1, Giovanni Sampaio Gonçalves2, Camino Fierro-Castro3
1Depatamento de Produção Animal. Faculdade de Ciências Agrárias e Tecnológicas, UNESP,
Campus de Dracena; Rod Cmte João Ribeiro de Barros, km 651- Dracena/SP, 17900-000,
Brasil. *E-mail: [email protected] 2SAA/APTA/Instituto de Pesca - Centro Avançado de Pesquisa Tecnológica do Agronegócio
do Pescado Continental, São José do Rio Preto /S,P 15025-970, Brasil. 3Departmento de Biología celular, Fisiología e Inmunología, Fac. Biociences, Universitat
Autònoma de Barcelona, Bellaterra, 08193 Spain.
The growth of Brazilian fish farming has led to an increase in the occurrence of diseases
of aquatic organisms, generating productive and economic losses. Paraller to this growth, the
aquaculture, in the 80’s, intensified studies to evaluate the use of immunostimulants, among
them the β- glucan, which has a powerful action on the non- specific immune system of fish,
promoting protection against many diseases. β-glucans are glucose polymers of fungal walls,
such as yeast Saccharomyces cerevisiae, with a potent immune action promoting serum proteins
concentration increase, such as lysozyme and complement system proteins, besides to stimulate
phagocytic activity of macrophages, leading to protection against many diseases. Thus, the aim
of the present study was to evaluate the β-glucan effects on the liver function, antioxidant
defenses and oxidative damages. For this purpose, tilapias were fed with a control diet for 90
days and an experimental diet with 10g kg-1 of β-glucan for 30, 60 and 90 days to verify the
existence of a prolonged effect of β-glucan over time. Fish from all treatments were sampled,
anesthetized in clove oil solution (1 g 10 L-1 of water), and blood was collected from 12 fish
(n=12) from each box (4 boxes per treatment) to evaluate glutathione (GSH), lipid peroxidation
measured by malondialdehyde (MDA) content and the hepatic somatic index (HSI). The results
indicated an increase of all variables after 30 days and values equal to the control diet at 60 and
90 days of treatment with the β-glucan diet, indicating an antioxidant adaptation to this diet.
We can only see significant decrease in hepatic GSH level of fish 30 days after stopping β-
glucan ingestion, which is most likely attributed to the decreased synthesis of glutathione
reductase. Regarding the MDA levels, no alterations were observed in liver compared to control
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
45
at 60 and 90 days with β-glucan diet and only when the fish were feed solely 30 days with β-
glucan show at significant increase of MDA content, indicating lipid peroxidation in the
analyzed tissues. The HSI like the rest of the markers, it is only shown modified at 30 days with
a slight increase. Our results demonstrate an adaptive response to β-glucan diet in fish, that diets
caused biochemical changes in liver cells, which in turn modulated enzymatic and non-
enzymatic antioxidants in Nile tilapia. It is concluded that β-glucan prolonged administration
do not impair antioxidant system, however other immunological variables should be evaluated
to verify the relationship between antioxidant and immune system of this compound.
Grants: The São Paulo Research Foundation - FAPESP – Process number: 2012/22016-
3.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
46
HEMATOLOGICAL STUDY OF THE EFFECTS OF PROLONGED
ADMINISTRATION OF Β-GLUCAN ON AN IMMUNITY, IN TILAPIA,
OREOCHROMIS NILOTICUS.
Camino Fierro-Castro1*, Ingrid Camargo dos Reis2, Basia Schlichting Moromizato2, Thais
Cristina de Oliveira2, Giovanni Sampaio Gonçalves3, Jaqueline Dalbello Biller2
1Departmento de Biología celular, Fisiología e Inmunología, Fac. Biociences, Universitat
Autònoma de Barcelona, Bellaterra, 08193 Spain. *E-mail:[email protected] 2Depatamento de Produção Animal. Faculdade de Ciências Agrárias e Tecnológicas, UNESP,
Campus de Dracena; Rod Cmte João Ribeiro de Barros, km 651- Dracena/SP, 17900-000,
Brasil. 3SAA/APTA/Instituto de Pesca - Centro Avançado de Pesquisa Tecnológica do Agronegócio
do Pescado Continental, São José do Rio Preto /S,P 15025-970, Brasil.
Is well known that immune-modulation of fish with β-glucans, using injection, dietary or
even immersion, has stimulating effects on both intestinal and systemic immunity. Although
the exact effects can be variable depending on fish species, administration route or time of
administration. The aim of this study was determining the best time line to administrate the β-
glucan diet studying the effects of dietary β-glucan supplementation (10g kg-1 of β-glucan), on
immune response, and antioxidant capacity of Tilapia, Oreochromis niloticus, under over time.
For this purpose, the tilapias were fed with a control diet for 90 days and an experimental
diet for 30, 60 and 90 days to verify the existence of a prolonged effect of β-glucan over time.
Fish from all treatments were sampled, anesthetized in clove oil solution (1 g 10 L-1 of water),
and blood was collected from 12 fish (n=12) from each box (4 boxes per treatment) to evaluate
hematological variables. Hemoglobin (HGB) content was assayed by cyanomethaemoglobin
determination. Wintrobe indexes such as mean corpuscular volume (MCV), corpuscular
hemoglobin means (CHM), and mean corpuscular hemoglobin concentration (MCHC) was
derived from the primary indexes. HGB concentrations and hematocrit did not differ among
treatments, but were within normal range for species, indicating that administration of β-glucan
does not alter fish energy production. Red blood cells (RBC) seems decreased after 60 and 90
days with β-glucan diet while MCV increased the same days. Given that the hemoglobin and
hematocrit values remain stable, this could be indicating the increase in the leukocyte
population involved in the immune defense system. This theory is reinforced with the study of
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
47
leukocyte respiratory activity Burst (NBT assay in blood) that show a significantly increases
after 90 days with a β-glucan diet. The knowledge of the hematological response under different
diets can be a useful tool for improve the administration of β-glucan and make it really effective.
Grants: The São Paulo Research Foundation - FAPESP – Process number:2012/22016-3.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
48
IN VITRO ACTION OF LACTIC ACID BACTERIA AGAINST BIOFILM-FORMING
LISTERIA MONOCYTOGENES
Juliana Sousa Bogéa1, Jucilene Sena dos Santos1, Luciane Manto2, Lucas Karpinski Risson3,
Franciele Maria Gottardo1, Lara Franco2, Laura Beatriz Rodrigues 1,2, Luciana Ruschel dos
Santos 1,2
1 Programa de Pós Graduação em Ciência e Tecnologia de Alimentos – PPCGTA.
Universidade de Passo Fundo, UPF.
2 Programa de Pós Graduação em Bioexperimentação – PPGBIOEXP,
Universidade de Passo Fundo, UPF
3 Bolsista PIBIC UPF, Curso de Biologia, Universidade de Passo Fundo, UPF
Listeria monocytogenes is a pathogenic microorganism that can be present in various
environments of the food industry and can cause serious economic and health damage, since
the bacterium causes listeriosis, a foodborne disease with high rates of mortality and
hospitalizations. The foods most associated with listeriosis are ready-to-eat products, especially
meat and dairy products. This fact is a major concern for the industry, since these products do
not necessarily suffer cooking or thermal inactivation prior to consumption. The presence of L.
monocytogenes in food production is associated with its ability to form biofilms, its high
tolerance to usual sanitizers, and its ability to multiply under refrigeration temperatures.
Biofilms are a set of organized living microorganisms surrounded by an extracellular polymeric
matrix that can be fixed and multiplied on any surface. Over time, they become permanent
reservoirs of pathogenic and deteriorating bacteria capable of resisting cleaning and disinfection
processes. The application of biological agents such as lactic acid bacteria (BALs) can be a
promising technique for controlling fixed pathogens on surfaces in the biofilms forms. The
application of BALs to neutralize the proliferation of other bacterial species on food contact
surfaces is based on the principle of competitive exclusion. In this technique, BALs with
aptitudes for the formation of protective biofilms play important roles in controlling
colonization by L. monocytogenes, since they can limit their proliferation through competition
for nutrients, the production of antimicrobial substances (bacteriocin), and the physical-
chemicals modifications of surfaces. In this work, the activity of BALs Bifidobacterium
animalis, Lactobacillus fermentum, L. plantarum, L. salivaris, and L. acidophilus was evaluated
against the formation of L. monocytogenes biofilms in polystyrene microplates. A strain of L.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
49
monocytogenes ATCC 7644 and another isolated in a stainless steel surface from a broilers
chicken slaughterhouse were used. This sample was subjected to tests of adhesion capacity in
microplates and resistance to sanitizers (sodium hypochlorite and hydrogen peroxide) in
previous works, showing adhesion capacity at all times (0, 4, 12, and 24 hours), temperatures (
Probiotics are live microorganisms that, when administered in adequate amounts, confer
a health benefit on the host. The main products that contain probiotics are dairy products,
however the consumption of these products may be limited by the number of individuals with
lactose intolerance and with cholesterol restricted diets. Thus, new products have been found
on the market, of non-dairy origin, which may have probiotic potential, such as Kombucha.
This beverage is prepared by fermenting sweetened black or green tea with a group of yeasts
and bacterias. The microbiological composition of the kombucha is variable, with some lactic
acid bacteria, such as Lactobacillus and Lactococcus, already detected in the beverages. These
represent a group of Gram-positive, catalase-negative microorganisms, with a characteristic of
lactic acid production, and are known for their probiotic properties. Some of these bacteria are
Enterococcus, Lactobacillus, Pediococcus, among others. The objective of this work was to
isolate, characterize and identify lactic acid bacteria in Kombuchas. Ten samples of Kombuchas
sold in Goiânia-GO, from five different brands and two flavors each, were collected. The
samples were diluted in series (10-1 to 10-3 and 10-6), then inoculated on Man Rogosa & Sharpe
(MRS) agar, selective for Lactobacillus sp., with surface plating method, incubated (37 ºC / 48
- 72 h) in anaerobiosis. Morphologically different isolates, similar to lactic acid bacteria, were
selected from Gram and catalase tests. From the microscopic analysis, a colony was selected
for molecular identification according to the Polymerase Chain Reaction technique using
specific primers from the V3-V4 region of the 16S rRNA gene. All samples showed growth of
lactic bacteria <106 CFU/mL, a population below what is necessary to claim probiotic
functionality in the product. Several authors have demonstrated the capacity of some lactic acid
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
58
bacteria with probiotic properties. During the production of the beverage, fermentation takes
place for 8 to 10 days, with production of metabolites, in which the viability of microorganisms
gradually decreases over the fermentation, due to the scarcity of oxygen and acidic conditions.
Still, the microbial environment can be affected by other factors, such as the ingredients
used in the product, temperature and storage time, as well as hygienic conditions of the
processing environment. According to Gram and catalase tests and morphological analysis, the
colonies showed characteristics consistent with lactic acid bacteria, being coco, catalase
negative and Gram positive. The molecular analysis identified the isolate as belonging to the
genus Pediococcus, of the species P. acidilactici. This species is associated with fermentative
processes of foods such as cereals, vegetables, meat products and fermented dairy products.
Recent studies have shown an important probiotic potential of P. acidilactici, due to its high
survival rate in gastric and duodenal conditions, inhibitory activity of pathogenic
microorganisms and production of antimicrobial peptide, pediocin. Therefore, commercial
Kombuchas can carry lactic acid bacteria of different morphologies, which are likely to have
probiotic potential such as Pediococcus acidilactici. However, further tests are needed to assess
the probiotic potential of the microorganism isolated from this beverage.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
59
LYOPHILIZED SYMBIOTIC MITIGATES MUCOSITIS INDUCED BY 5-
FLUOROURACIL
Bruna Savassi1, Bárbara F. Cordeiro1, Emiliano R. Oliveira1, Sara H. Silva1, Giovanna Belo1,
Erick A. Esmerino2, Ramon S. Rocha2, Erick A. Esmerino2, Monica Q. Freitas2, Marcia C.
Silva3, Adriano G. Cruz3*, Kátia Duarte Vital4, Simone O.A. Fernandes4, Valbert N.
Cardoso4, Alfonso Gala-Garcia1, Vasco Azevedo1, Fillipe Luiz R. do Carmo1,5
1 Universidade Federal de Minas Gerais (UFMG), Instituto de Ciências Biológicas (ICB)
2 Universidade Federal Fluminense (UFF), Faculdade de Veterinária 3 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos, email:[email protected] 4 Universidade Federal de Minas Gerais (UFMG), Departamento de Análises Clínicas e
Toxicológicas 5 Science et Technologie du Lait et de l’Oeuf, Institut National de la Recherche Agronomique,
Agrocampus Ouest, Rennes, France.
Mucositis is a common side effect of cytotoxic chemotherapy with 5-FU, characterized
by mucosal injury, inflammation, diarrhea, and weight loss. Previous studies show promising
results of probiotic strains associated with prebiotics as adjuvant treatment for mucositis. We
developed a symbiotic product lyophilized, containing skimmed milk supplemented with Whey
Protein Isolate (WPI) fermented by Lactobacillus (L.) casei BL23, L. plantarum B7 and L.
rhamnosus D1 and added of fructooligosaccharides (FOS). In the 5-Fluorouracil mucositis mice
model, the symbiotic lyophilized formulation reduced (p <0.05) the animals' weight loss,
10.15% symbiotic group to 13.85 % inflamed group. Moreover, symbiotic treatment
significantly reduced (p <0.05) intestinal permeability (0.04%) quantify by radioactivity of
99mTc-DTPA in the blood compared to inflamed control group (0.13%). Finally, histological
damage caused by 5-FU-induced mucositis was measured and exhibited a reduced score of
inflammation in duodenum, ileum and colon in mice treated with symbiotic formulation (p
<0.05). In addition, symbiotic treatment was able to reducing (p <0.05). the production of the
pro-inflammatory cytokine IL-1β in spleen (205 pg/mL) compared to inflamed group
(463pg/mL). Therefore, it was possible to conclude that the symbiotic product developed in
this work represents a promising adjuvant treatment of mucositis.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
60
METABOLIC PROFILING OF PROBIOTIC LOW-SODIUM PRATO CHEESE
WITH FLAVOR ENHANCERS: USEFULNESS OF NMR SPECTROSCOPY AND
CHEMOMETRIC TOOLS
Celso F. Balthazar1, Jonas T. Guimarães1, Ramon S. Rocha1,2, Roberto P.C. Neto3, Erick A.
Esmerino1, Hugo L. A. Silva1, Elenilson G. Alves Filho4, Tatiana C. Pimentel5, Sueli
Rodrigues4, Edyr S. Brito6, Maria Inês B. Tavares3, Daniel Granato7, Marcia Cristina Silva2,
Renata S.L. Raices2, Mõnica Q. Freitas1, Adriano G. Cruz2*
1 Universidade Federal Fluminense (UFF), Faculdade de Veterinária
2 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos, email:[email protected] 3 Universidade Federal do Rio de Janeiro (UFRJ), Instituto de Macromoléculas Professora
Eloisa Mano (IMA) 4 Universidade Federal do Ceará (UFC), Departamento de Engenharia de Alimentos 5 Instituto Federal de Educação, Ciência e Tecnologia do Paraná (IFPR), Paranavaí
6 Embrapa Agroindustria Tropical, Fortaleza-CE, Brazil. 7 Food Processing and Quality, Production Systems Unit, Natural Resources Institute Finland
(Luke),
The metabolic profiling of probiotic low-sodium Prato cheeses added with flavor
enhancers (arginine, yeast extract, and oregano extract) along the ripening time (1, 30, 60 days)
using NMR spectroscopy and chemometric tools (PCA and PLS-DA) was evaluated. Aromatic
and aliphatic structures and lactic acid were the most dominant compounds regardless of the
flavor enhancer added. The addition of flavor enhancers decreased the lactic acid content and
increased the content of leucine, isoleucine, and valine, fatty acids, tyrosine, phenylalanine,
histidine, benzoic acid, and formic acid. Principal component analysis (PCA) differentiated
cheese samples based on the contents of lactic, acetic, formic, benzoic, and fatty acids, valine,
leucine, isoleucine, tyrosine, phenylalanine, and histidine. Partial least square discriminant
analysis (PLSDA) classified the cheese types based on composition, as well as highlighted the
compounds variability according to the ripening time. NMR spectroscopy and chemometrics
may be an effective approach to investigate the metabolic profiling of probiotic low-sodium
Prato cheese with the addition of flavor enhancers.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
61
MICROPARTÍCULAS DE KEFIR E EXERCÍCIO INTENSO EM MODELO
ANIMAL
ANA LUIZA COUTINHO FAVILLA, MILENA DE CÁSSIA DOS SANTOS DA SILVA,
LUAN RIBEIRO E ANNA PAOLA PIERUCCI
UNIVERSIDADE FEDERAL DO RIO DE JANEIRO
CENTRO DE CIÊNCIAS DA SAÚDE (CCS)
INSTITUTO DE NUTRIÇÃO JOSUÉ DE CASTRO
LABORATÓRIO DE DESENVOLVIMENTO DE ALIMENTOS PARA FINS ESPECIAIS
E EDUCACIONAIS
A habilidade da microbiota de metabolizar energia, de modular o sistema imune e de
alterar parede intestinal, atua de forma significante na saúde de atletas e na performance durante
a prática da atividade física. A suplementação de kefir em modelos de exercício já foi associada
a redução a de níveis de proteína C reativa, regulação negativa da expressão de mediadores pró-
inflamatórios, como IL-6 e IFN-gama, melhora da performance e redução da fadiga. A secagem
e a encapsulação spray drying pode otimizar a viabilidade, a funcionalidade e a resistência,
além de facilitar e baratear os custos de transporte a estocagem de probióticos. Nesse estudo, a
intensidade de protocolo de exercício e os efeitos do consumo de micropartículas de kefir em
ratos expostos a atividade física intensa foi estudada através do dano celular e de lactato. Ratos
Wistar machos (N=15), com 8 semanas foram divididos aleatoriamente em 4 grupos. A um
grupo (n=5 para cada) foi administrado kefir encapsulado com maltodextrina reconstituído
(KE), a outro grupo controle leite desnatado (LD) e ao terceiro grupo foi administrado leite
fermentado de kefir (KF), por gavagem intragástrica. Todos os grupos foram exposto ao
exercício high intensity interval training (HIIT) em esteira rolante com malha de choque. Foram
quantificados lactato, AST, ALT e concentração de AGCC. Quanto ao teor de lactato, não
houve diferença em nenhum dos momentos experimentais em nenhum dos grupos. Entretanto,
os valores absolutos encontrados foram mais elevados do que os descritos na literatura para
modelo de exercício em animais, sugerindo uma elevada intensidade desse modelo. Os três
grupos mostraram aumento nos valores de ALT, após duas semanas de administração de kefir.
O grupo LD tinha o menor valor no momento basal e foi o que apresentou o maior valor em
após duas semanas de administração de kefir, sendo esta diferença significativa (P-Valor =
0,0001), sugerindo uma maior lesão do músculo esquelético, mas sabe-se que essa alteração
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
62
pode ser fisiológica. Na semana de suplementação aumento de ALT no grupo LD, pode ser
explicado pela propriedade inflamatória do leite de vaca. Houve aumento de AST nos grupos
LD (significativo, P-valor= 0,0054) e LF do após a introdução dos leites. AST tende a aumentar
após exercício exaustivo sem a utilização de probióticos e no grupo KE, houve redução
significativa após a combinação com o exercício intenso. Foram detectados ácido acético, ácido
butírico e ácido propiônico. Os resultados não mostraram diferença significativa para nenhum
ácido testado em nenhuma das suplementações. Os dados revelam primariamente uma elevada
intensidade do exercício HIIT, mas não alteração da AGCC pelo kefir. O kefir com material
encapsulante não promoveu resultado comparativamente melhor nos parâmetros testados. O
leite de vaca, utilizado como controle, provocou aumento dos marcadores analisados,
mostrando potencial inflamatório deste. Com ajustes, o modelo pode ser adequado a estudos
subsequentes que irão avaliar a alteração da permeabilidade intestinal e da composição da
microbiota, mediante exercício extenuante.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
63
MICROWAVE TREATMENT CHANGES THE TRIDIMENSIONAL STRUCTURE
OF INSOLUBLE FIBERS AND IMPROVES THEIR FERMENTATION PROFILE BY
HUMAN GUT MICROBIOTA
Thaisa Junglesa; Xiaowei Zhanga, Bruce Hamakera ;Lucimara Cordeirob;
a Whistler Center for Carbohydrate Research and Department of Food Science, Purdue
University, West Lafayette, IN 47907, USA. b Department of Biochemistry, Federal University of Parana, Curitiba- Brazil.
Abstract: Most of the insoluble dietary fibers are known to be poorly fermented by the
human gut microbiota. Since microwave (MW) treatment is capable to enhance the
susceptibility of insoluble materials to enzymatic hydrolysis we tested if MW would enhance
insoluble dietary fibers fermentability in an in vitro gut fermentation model. As expected, 24
hours in vitro fecal fermentation of insoluble xylan, arabinoxylan, and arabinan did not increase
the production of gas nor short-chain fatty acids (SCFA) if compared to the blank, indicating
low fermentability. However, after microwave treatment (800W, 180oC, 30 min) significant
increase in fermentation metabolites produced including the main SCFA (acetate, butyrate and
propionate) could be observed for all tested dietary fibers fermented after MW. Thus, MW
irradiation was capable to increase the fermentation profile of the tested insoluble dietary fibers.
The increased fermentability of MW dietary fibers was partially explained by shifts in the
tridimensional structure, which were observed through scanning electron microscopy. Finally,
16S rRNA gene sequencing of samples indicate that distinct bacterial groups were stimulated
according to the tested dietary fiber type. These results allow for the design of insoluble but yet
fermentable prebiotic dietary fiber of health significance using microwave irradiation.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
64
OHMIC HEATING OF PROBIOTIC FERMENTED MILK : ANTI-CANCER
ACTIVITY, FATTY ACIDS PROFILE, VOLATILE COMPOUNDS, AND SENSORY
ATTRIBUTES
Amanda B. Silva1, Hugo Scudino1, Jonas T. Guimarães1, Ramon S. Rocha1,2, Gustavo L.P.A.
Ramos1,2, Felipe S. Lemos5, Tatiana C. Pimentel3, Daniel Granato4, Felipe L. Oliveira5,
Celso F. Balthazar1, Jonas T. Guimarães1, Ramon S. Rocha1,2, Roberto P.C. Neto3, Erick A.
Esmerino1, Hugo L. A. Silva1, Elenilson G. Alves Filho4, Tatiana C. Pimentel5, Sueli
Rodrigues4, Edyr S. Brito6, Maria Inês B. Tavares3, Daniel Granato7, Marcia Cristina Silva2,
Renata S.L. Raices2, Mõnica Q. Freitas1, Adriano G. Cruz2
1 Universidade Federal Fluminense (UFF), Faculdade de Veterinária
2 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos, e-mail: [email protected] 3 Universidade Federal do Rio de Janeiro (UFRJ),
Instituto de Macromoléculas Professora Eloisa Mano (IMA) 4 Universidade Federal do Ceará (UFC), Departamento de Engenharia de Alimentos 5 Instituto Federal de Educação, Ciência e Tecnologia do Paraná (IFPR), Paranavaí
6 Embrapa Agroindustria Tropical, Fortaleza-CE, Brazil. 7 Food Processing and Quality, Production Systems Unit,
Natural Resources Institute Finland (Luke),
This study aimed to evaluate the impact of the addition of L. casei 01 as a probiotic culture
on the metabolic profile of Minas Frescal cheese using NMR (Nuclear Magnetic Resonance)
spectroscopy and chemometric technique (Principal Component Analysis, PCA). In addition,
the volatile compounds was also analyzed. Lactose (α-lactose and β-lactose), fatty acids
(unsaturated and saturated), citric acid, tryptophan, and benzoic acid were observed as main
compounds. Probiotic cheese was characterized by the highest concentration of tryptophan and
presented a higher number of volatile acids. The control cheese was characterized by the
highest concentration of benzoic acid, and fatty acids, resulting in a higher number of volatile
alcohols and esters. No differences were observed for α-lactose, β-lactose, and citric acid
contents. A clear separation of probiotic and control Minas frescal cheese was observed in
PCA of the data of the 1H NMR spectra, demonstrating that the addition of probiotic culture
altered the metabolic profile of Minas Frescal cheese. Overall, the findings suggest that the
addition of probiotic culture promoted the proteolysis in the fresh cheeses, decreased the
lipolysis, and altered the volatile compounds. Furthermore, NMR spectroscopy and
chemometrics tools could be used to differentiate probiotic and conventional cheeses
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
68
PROBIOTIC PRATO CHEESE ENHANCED THE COLONIC BARRIER
INTEGRITY IN IN MICE WITH ULCERATIVE COLITIS
Bárbara F. Cordeiro1, Emiliano R. Oliveira1, Sara H. Silva1, Giovanna Belo1, Bruna Savassi1,
Alessandra Figueiroa1, Erick A. Esmerino2, Ramon S. Rocha2, Monica Q. Freitas2, Marcia C.
Silva3, Adriano G. Cruz3*, Fillipe Luiz R. Carmo1,4, Vasco Azevedo1
1 Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais (UFMG) 2 Universidade Federal Fluminense (UFF), Faculdade de Veterinária
3 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos, e-mail: [email protected] 4Science et Technologie du Lait et de l’Oeuf, Institut National de la Recherche Agronomique
The role of experimental probiotic Prato cheese containing Lactobacillus casei 01 in the
prevention of dextran sodium sulphate (DSS)-induced ulcerative colitis in mice was evaluated.
For the DSS in vivo model, mice were divided into six groups. The groups 1–3 represented
noninflamed groups and groups 4–6 received a DSS (2%) solution. Mice from groups 1 and 4
intragastrically received 500 μL of phosphate-buffered saline (control groups), while mice from
groups 2 and 5 intragastrically received 500 μL of conventional Prato cheese (control groups).
Finally, mice from group 3 and 6 intragastrically received 500 μL of Prato probiotic cheese
containing L. casei 01 (9.47 log cfu mL−1). The groups treated with probiotic Prato cheese
exhibited reduced weight loss caused by the consumption of DSS solution. However, the
probiotic cheese failed to reduce the inflammation scores in DSS-induced colitis group. Overall,
probiotic Prato cheese was not effective to ameliorate the symptoms of DSS-induced colitis.
The results showed that the probiotic cheese enhanced the colonic barrier integrity index and
stimulated muc2 gene expression in healthy animals. However, the experimental probiotic
cheese failed to reduce the macroscopic and microscopic inflammatory scores of DSS-induced
colitis. Overall, probiotic Prato cheese contains L. casei 01 was not effective to ameliorate the
symptoms of DSS-induced colitis.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
69
ANTIHYPERTENSIVE EFFECT OF A PROBIOTIC BEVERAGE IN
SPONTANEOUSLY HYPERTENSIVE RATS
Alessandra Figueiroa1, Bárbara F. Cordeiro1, Emiliano R. Oliveira1, Sara H. Silva1, Giovanna
Belo1, Erick A. Esmerino2, Ramon S. Rocha2, Monica Q. Freitas2, Marcia C. Silva3, Adriano
G. Cruz3, Alfonso Gala-Garcia1, Vasco Azevedo1, Fillipe Luiz R. do Carmo1,4
1, Universidade Federal de Minas Gerais (UFMG), Instituto de Ciências Biológicas (ICB)
2 Universidade Federal Fluminense (UFF), Faculdade de Veterinária 3 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos 4Science et Technologie du Lait et de l’Oeuf, Institut National de la Recherche Agronomique,
Agrocampus Ouest, Rennes, France.
The Lactobacillus (L.) delbrueckii subsp. lactis CNRZ327 has shown promising results
in experimental models of colitis induced by chemical agents, which concurrently promote
ACE inhibitory activity. To investigate the systemic hypotensive effect of L. delbrueckii
CNRZ327, this strain was evaluated in an in vivo assay using SHR rats (spontaneously
hypertensive rats). We developed a probiotic beverage fermented by L. delbrueckii CNRZ327
and supplemented with Whey Protein Isolated (WPI) 30%. CFU counting of L. delbrueckii
CNRZ327 in probiotic beverage is 2,25 x 109 CFU/mL. Probiotic beverage exhibited higher
Angiotensin-converting-enzyme (ACE)-inhibition activity (78,9% ± 0,11) and the antioxidant
activity (86,5 % ± 0,26). Finally, in the in vivo model, probiotic beverage showed a significant
reduction (p <0.05) of systolic blood pressure (177,043 ± 3.80 mmHg) for the 1st experimental
day to final experimental day 14th (139.85 ± 7.83 mmHg). It is possible that the presence of the
angiotensin-converting enzyme has generated this effect, which makes the probiotic beverage
fermented by L. delbrueckii subsp. lactis CNRZ327 and supplemented with WPI promising in
the prevention and / or treatment of arterial hypertension.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
70
LYOPHILIZED SYMBIOTIC MITIGATES MUCOSITIS INDUCED BY 5-
FLUOROURACIL
Bruna Savassi1, Bárbara F. Cordeiro1, Emiliano R. Oliveira1, Sara H. Silva1, Giovanna Belo1,
Erick A. Esmerino2, Ramon S. Rocha2, Monica Q. Freitas2, Marcia C. Silva3, Adriano G.
Cruz3, Kátia Duarte Vital4, Simone O.A. Fernandes4, Valbert N. Cardoso4, Alfonso Gala-
Garcia1, Vasco Azevedo1, Fillipe Luiz R. do Carmo1,5
1 Universidade Federal de Minas Gerais (UFMG), Instituto de Ciências Biológicas (ICB)
2 Universidade Federal Fluminense (UFF), Faculdade de Veterinária 3 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos 4 Universidade Federal de Minas Gerais (UFMG),
Departamento de Análises Clínicas e Toxicológicas 5 Science et Technologie du Lait et de l’Oeuf, Institut National de la Recherche Agronomique,
Agrocampus Ouest, Rennes, France.
Mucositis is a common side effect of cytotoxic chemotherapy with 5-FU, characterized
by mucosal injury, inflammation, diarrhea, and weight loss. Previous studies show promising
results of probiotic strains associated with prebiotics as adjuvant treatment for mucositis. We
developed a symbiotic product lyophilized, containing skimmed milk supplemented with Whey
Protein Isolate (WPI) fermented by Lactobacillus (L.) casei BL23, L. plantarum B7 and L.
rhamnosus D1 and added of fructooligosaccharides (FOS). In the 5-Fluorouracil mucositis mice
model, the symbiotic lyophilized formulation reduced (p <0.05) the animals' weight loss,
10.15% symbiotic group to 13.85 % inflamed group. Moreover, symbiotic treatment
significantly reduced (p <0.05) intestinal permeability (0.04%) quantify by radioactivity of
99mTc-DTPA in the blood compared to inflamed control group (0.13%). Finally, histological
damage caused by 5-FU-induced mucositis was measured and exhibited a reduced score of
inflammation in duodenum, ileum and colon in mice treated with symbiotic formulation (p
<0.05). In addition, symbiotic treatment was able to reducing (p <0.05). the production of the
pro-inflammatory cytokine IL-1β in spleen (205 pg/mL) compared to inflamed group
(463pg/mL). Therefore, it was possible to conclude that the symbiotic product developed in
this work represents a promising adjuvant treatment of mucositis.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
71
PROBIOTICOS PARA PETS: MICROENCAPSULAÇÃO E SIMULAÇÃO DAS
0,020% de sal de cura e probiótico microencapsulado com alginato de sódio através do método
de extrusão ( > 6 log UFC/g). As copas foram maturadas em estufa BOD na temperatura de 15-
18 ºC por 30 dias. Posteriormente, foram analisadas possíveis alterações nas características
físico-químicas e sensoriais, bem como a viabilidade da cultura no produto cárneo. Todas as
amostras apresentaram-se dentro da legislação brasileira para os atributos de umidade, proteína
e lipídios. Os salames no início apresentaram valores de pH entre 6,19 e 6,30 e durante o tempo
de fermentação os valores diminuíram até o dia 15, quando voltaram a aumentar. Ao final do
período de maturação, a perda de peso das copas foram de 40,48; 44,6 e 55,77% para amostras
C, BB1 e BB2, respectivamente. Em relação a cor houve diferença significativa entre os
tratamentos sendo que os parâmetros L* e b* apresentaram redução e a* apresentou aumento
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
74
ao final de 30 dias. A adição de probióticos afetou os valores do TBARS e houve diferença
significativa entre a amostra controle e as com a adição de BB12 durante o período de
maturação. A amostra controle apresentou maiores valores de TBARS nos dias 0, 15 e 30 em
comparação às amostras com a adição de BB12. A viabilidade dos probióticos
microencapsulados apresentaram-se viáveis após o período de maturação de 30 dias no
tratamento BB2. Não houve diferença significativa entre as amostras para a análise sensorial
sendo então, possível afirmar que a adição de BB12 é uma alternativa para se obter um produto
cárneo com todos os benefícios de alimentos funcionais.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
75
PROPRIEDADES FÍSICO-QUÍMICAS E CARGA MICROBIOLÓGICA EM
MICROPARTÍCULAS DE KEFIR
MILENA DE CÁSSIA DOS SANTOS DA SILVA, ANA LUIZA FAVILLA, MARIA
CLAUDIA RODRIGUES E ANNA PAOLA PIERUCCI
UNIVERSIDADE FEDERAL DO RIO DE JANEIRO
CENTRO DE CIÊNCIAS DA SAÚDE (CCS)
INSTITUTO DE NUTRIÇÃO JOSUÉ DE CASTRO
LABORATÓRIO DE DESENVOLVIMENTO DE ALIMENTOS PARA FINS ESPECIAIS
E EDUCACIONAIS
A microencapsulação é um processo pelo qual os materiais bioativos são revestidos
com outros materiais de proteção ou suas misturas e a secagem por spray é um processo
amplamente utilizado para a preservação de alimentos, garantindo uma vida útil estável e
prolongada, reduzindo custos de transporte e facilitando a comercialização. Nos últimos anos,
muitos estudos foram conduzidos sobre microencapsulação para a preservação de
microrganismos probióticos durante o armazenamento e processamento de alimentos. Visando
verificar e quantificar o rendimento, as propriedades físico-químicas e as propriedades
microbiológicas do kefir foram elaboradas micropartículas dessa substância, empregando
“Spray Dryer”. A fermentação dos grãos em leite desnatado ocorreu a 25°C/24 h, com
maturação de 4°C/24 h. Foram executados processos de microencapsulamento sem matriz
encapsulante (SM) (130 a 180°C), com maltodextrina (MD) 1:1 e com Capsul® (CAP) 1:1 (140
a 160°C). Não houve diferença significativa entre os rendimentos (40,6 e 54,3 %) dos processos
de microencapsulamento. O processo fermentativo provocou um importante aumento na acidez
do leite (de 0,12 a 1,03g / 100g) e redução acentuada do pH após a fermentação (de 6,67 para
4,04). O teor de umidade das micropartículas de kefir variou nas três amostras em triplicata
(SM - 16,92 a 18,55%; MD – 4,40 a 5,07%; CAP – 5,34 a 6,50%) e a atividade de água (Aw)
das micropartículas de kefir ficou entre 0,224 e 0,315, situando-se na faixa adequada para
produtos desidratados. A contagem de bactérias viáveis (Log UFC/g em base úmida) foi de
6,11 para fungos filamentosos e leveduras; 9,64 para bactérias ácido-láticas; 10,20 para
lactobacilos; 10,00 para mesófilos e 7,78 para bactérias ácido-acéticas. As bactérias ácido-
láticas e lactobacilos permaneceram viáveis em todas as temperaturas de secagem (9,88 e 9,87
a 130 °C, e 3,79 e 4,61 à temperatura de 180 °C, respectivamente). Os fungos filamentosos e
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
76
leveduras e as bactérias mesófilas mostraram-se mais sensíveis ao aumento da temperatura, a
partir de 150°C para fungos filamentoso em todas as amostras e para bactérias mesófilas nas
amostras com matriz encapsulante e a partir de 170°C para bactérias mesófilas sem matriz
encapsulante. Bactérias ácido-acéticas demonstraram ser as mais resistentes dentre os grupos
de microrganismos analisados, apresentando redução na contagem apenas nas duas
temperaturas mais elevadas, 6,56 a 170°C e 4,73 a 180°C. Portanto, observa-se que o
rendimento, a análise das propriedades físico-químicas e contagem microbiana foram
satisfatórios e corroboraram com os estudos encontrados na literatura. Além disso, processo
forneceu condições apropriadas à preservação da carga microbiológica nas micropartículas de
kefir.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
77
EVALUATION OF ANTIOXIDANT RESPONSE AFTER PROLONGED
ADMINISTRATION OF Β-GLUCAN ON TILAPIA, OREOCHROMIS NILOTICUS.
Jaqueline Dalbello Biller1*, Ingrid Camargo dos Reis1, Eduardo Antônio Oliveira Cavalheiro1,
Victor Negri Libório1, Giovanni Sampaio Gonçalves2, Camino Fierro-Castro3
1Depatamento de Produção Animal. Faculdade de Ciências Agrárias e Tecnológicas, UNESP,
Campus de Dracena; Rod Cmte João Ribeiro de Barros, km 651- Dracena/SP, 17900-000,
Brasil. *E-mail: [email protected] 2SAA/APTA/Instituto de Pesca - Centro Avançado de Pesquisa Tecnológica do Agronegócio
do Pescado Continental, São José do Rio Preto /S,P 15025-970, Brasil. 3Departmento de Biología celular, Fisiología e Inmunología, Fac. Biociences, Universitat
Autònoma de Barcelona, Bellaterra, 08193 Spain.
The growth of Brazilian fish farming has led to an increase in the occurrence of diseases
of aquatic organisms, generating productive and economic losses. Paraller to this growth, the
aquaculture, in the 80’s, intensified studies to evaluate the use of immunostimulants, among
them the β- glucan, which has a powerful action on the non- specific immune system of fish,
promoting protection against many diseases. β-glucans are glucose polymers of fungal walls,
such as yeast Saccharomyces cerevisiae, with a potent immune action promoting serum proteins
concentration increase, such as lysozyme and complement system proteins, besides to stimulate
phagocytic activity of macrophages, leading to protection against many diseases. Thus, the aim
of the present study was to evaluate the β-glucan effects on the liver function, antioxidant
defenses and oxidative damages. For this purpose, tilapias were fed with a control diet for 90
days and an experimental diet with 10g kg-1 of β-glucan for 30, 60 and 90 days to verify the
existence of a prolonged effect of β-glucan over time. Fish from all treatments were sampled,
anesthetized in clove oil solution (1 g 10 L-1 of water), and blood was collected from 12 fish
(n=12) from each box (4 boxes per treatment) to evaluate glutathione (GSH), lipid peroxidation
measured by malondialdehyde (MDA) content and the hepatic somatic index (HSI). The results
indicated an increase of all variables after 30 days and values equal to the control diet at 60 and
90 days of treatment with the β-glucan diet, indicating an antioxidant adaptation to this diet.
We can only see significant decrease in hepatic GSH level of fish 30 days after stopping β-
glucan ingestion, which is most likely attributed to the decreased synthesis of glutathione
reductase. Regarding the MDA levels, no alterations were observed in liver compared to control
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
78
at 60 and 90 days with β-glucan diet and only when the fish were feed solely 30 days with β-
glucan show at significant increase of MDA content, indicating lipid peroxidation in the
analyzed tissues. The HSI like the rest of the markers, it is only shown modified at 30 days with
a slight increase. Our results demonstrate an adaptive response to β-glucan diet in fish, that diets
caused biochemical changes in liver cells, which in turn modulated enzymatic and non-
enzymatic antioxidants in Nile tilapia. It is concluded that β-glucan prolonged administration
do not impair antioxidant system, however other immunological variables should be evaluated
to verify the relationship between antioxidant and immune system of this compound.
Grants: The São Paulo Research Foundation - FAPESP – Process number: 2012/22016-
3.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
85
PARAPROBIÓTICOS: RELEVÂNCIA DO AQUECIMENTO ÔHMICO COMO
TECNOLOGIA DE OBTENÇÃO
Cássia P. Barros1, Roberto P.S. Pires2, Ramon S. Rocha1,2, Ramon Silva 1,2, Jonas T.
Guimarães1, Yuri K.D. Abud3, Celso Sant'Anna3, Luiz Dione B. Melo2, Maria Carmela K. H.
Duarte1, Erick A. Esmerino1, Márcia C. Silva2, Beatriz M Ferreira4, Anderson S. Sant’Ana4,
Monica Q. Freitas1, Adriano G. Cruz2
1Universidade Federal Fluminense (UFF),
Faculdade de Medicina Veterinária, Niterói, 24230-340, Rio de Janeiro, Brasil 2Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos, Rio de Janeiro, 20270-021, Brasil 3Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO),
Duque de Caxias, 25250-020, Rio de Janeiro, Brasil 4 Universidade de Campinas (UNICAMP), Faculdade de Engenharia de Alimentos (FEA),
Campinas, SP, Brasil
Parabrobióticos são conhecidos como probióticos inativados (mortos) que, conforme
observado em pesquisas, também podem fornecer benefícios à saúde ao exercer efeitos
imunomoduladores no hospedeiro. Embora sejam usualmente produzidos pelo tratamento
térmico, tecnologias não-convencionais apresentam-se como uma alternativa interessante para
sua obtenção. Objetivou-se com o presente estudo avaliar o efeito do aquecimento ôhmico (AO)
como método de inativação nas propriedades celulares e morfológicas das cepas probióticas
comerciais Lactobacillus acidophilus LA 05 (LA), Lactobacillus casei 01 (LC) e
Bifidobacterium animalis Bb 12 (BA) sob diferentes gradientes de campo elétrico (4, 8 e 12
V/cm a 60 Hz), gerando paraprobióticos. AO foi comparado com o tratamento térmico
convencional (CONV) a fim de determinar a condição mais adequada para produção de
paraprobióticos, por meio da culturabilidade como metodologia convencional, citometria de
fluxo (CF) e microscopia eletrônica de varredura (MEV). O tratamento CONV proporcionou
perda da culturabilidade e, portanto, inativação de cada cepa nas seguintes condições: L.
acidophilus - 95°C/ 5 minutos, L. casei e B. animalis - 95°C/ 7 minutos. Esse mesmo perfil de
tempo e temperatura foi utilizado no AO para comparar os tratamentos. Na CF foram
observados diferentes graus de danos no potencial e integridade da membrana celular induzidos
pelos tratamentos. Entretanto, foi constatado que o AO na condição 8 V/cm, 95°C/ 7 minutos a
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
86
60 Hz causou menor grau de injúria ao apresentar menor percentual de células com membranas
permeabilizadas (78,1%, 78,9% e 88,9% de LC, BA e LA, respectivamente, p < 0,05)
comparado ao tratamento CONV (88,1%, 88,1% e 89,5% de LC, BA e LA, respectivamente, p
< 0,05), revelando assim, danos térmicos mais brandos. Alterações morfológicas como
rugosidade na superfície celular, extravasamento de conteúdo citoplasmático e detritos
celulares foram observadas na MEV. A CF em consonância com a MEV forneceu informações
complementares a culturabilidade que foram extremamente úteis para escolha da melhor
condição de inativação por AO. Os resultados obtidos sugerem que o AO é uma tecnologia
adequada para produção de paraprobióticos.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
87
WHEY DRINK DE UVA PARAPROBIÓTICO DE LACTOBACILLUS CASEI
OBTIDO POR AQUECIMENTO ÔHMICO É EFICAZ NO CONTROLE DA
GLICEMIA PÓS-PRANDIAL EM ADULTOS SAUDÁVEIS
Cássia P. Barros1, Roberto P.S. Pires2, Ramon S. Rocha1,2, Ramon Silva1,2, Jonas T,
Guimarães1, Elisa L. Venâncio3, Itallo Collopy Junior3, Paula M.C. Maciel3, Beatriz M.
Ferreira4, Anderson S. Sant’Ana4, Maria Carmela K. H. Duarte1, Marcia C. Silva1, Mônica Q.
Freitas1, Adriano G. Cruz2
1Universidade Federal Fluminense (UFF), Faculdade de Medicina Veterinária, 24230-340,
Niterói, Rio de Janeiro, Brasil 2Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Departamento de Alimentos, 20270-021, Rio de Janeiro, Brasil 3 Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ),
Curso de Farmácia, 21715-000, Realengo, RJ, Brasil 4 Universidade de Campinas (UNICAMP), Faculdade de Engenharia de Alimentos (FEA),
Campinas, SP, Brasil
O consumo de probióticos lácteos está associado a diversos benefícios à saúde, como
atividade anti-hiperglicêmica, ao auxiliar na redução da glicemia pós-prandial. Entretanto,
conforme observado em pesquisas, probióticos inativados denominados “paraprobióticos”
também conferem benefícios ao hospedeiro, além de proporcionar maior segurança e
conveniência para manuseio industrial. Porém, até o momento, não existem estudos envolvendo
a atividade anti-hiperglicêmica de paraprobióticos obtidos por metodologias emergentes. O
objetivo do presente estudo foi investigar o efeito da ingestão do whey drink de uva
paraprobiótico de Lactobacillus casei 01 inativado por aquecimento ôhmico (8 V/cm, 95°C/ 7
minutos a 60 Hz) na atividade anti-hiperglicêmica in vitro e in vivo comparado ao whey drink
de uva probiótico (7-8 log UFC/ml). A atividade anti-hiperglicêmica in vitro foi analisada pela
inibição da atividade das enzimas α-glucosidase e α-amilase, enquanto o estudo in vivo foi
conduzido com indivíduos saudáveis (n = 15, sendo 3 homens e 12 mulheres, 23-46 anos),
que tiveram a glicemia pós-prandial mensurada antes (jejum) e após o consumo dos seguintes
tratamentos: pão + água (controle), pão + whey drink de uva probiótico e pão + whey drink de
uva paraprobiótico. No estudo in vitro, observou-se a inibição da atividade das enzimas α-
glucosidase e α-amilase de forma semelhante no whey drink probiótico e paraprobiótico em
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
88
relação ao controle (51,2% e 43,3% e 51,8% e 44,3%, respectivamente, p > 0,05). Enquanto no
estudo in vivo, o consumo do pão isolado (controle) resultou em menor aumento da glicemia
(36 mg/dl, p < 0,05) quando comparado ao whey drink paraprobiótico e probiótico (46 mg/dl e
49 mg/dl, respectivamente p > 0,05). No entanto, os valores dos demais índices glicêmicos
analisados, como porcentagem incremental de glicose (paraprobiótico e probiótico, 50,3% e
54,7%, respectivamente, p > 0,05) e a redução da resposta glicêmica (paraprobiótico e
probiótico, 13,1 mg/dl e 13,4 mg/ dl, respectivamente, p > 0,05) se mantiveram análogos ao
controle (39,8% e 12,4 mg/dl, p > 0,05) apesar do whey drink de uva possuir açúcares inerentes
a sua composição, o que demonstra capacidade de controle da taxa glicêmica. Tais resultados
são congruentes com o estudo in vitro, sugerindo uma boa correlação das bebidas
paraprobiótica e probiótica na atividade anti-hiperglicêmica. Sendo assim, conclui-se que o
whey drink de uva paraprobiótico representa uma alternativa tão eficaz quanto o probiótico na
redução da resposta glicêmica pós-prandial em indivíduos saudáveis.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
89
CARACTERIZAÇÃO SENSORIAL DE GELADO COMESTÍVEL Á BASE DE
EXTRATO AQUOSO DE SOJA, KEFIR E JABOTICABA
Giovana Maria Navarro de Mendonça; Eliane Vale da Silva; Mariela Zucolo; Daniela
Cardoso Umbelino Cavallini.
A busca por alimentos que promovam a saúde tem motivado a diversificação do mercado
de alimentos funcionais. Os gelados comestíveis, conhecidos popularmente como sorvetes,
apresentam composição variada, incluindo açúcares, proteínas, polpas, sucos e outras matérias-
primas, permitindo, desta forma, a obtenção de diferentes formulações. Nesse contexto, o
objetivo deste estudo foi o desenvolvimento de um gelado comestível potencialmente
funcional, a partir do extrato aquoso de soja (EHS), kefir e casca de jaboticaba desidratada.
Foram produzidos cinco gelados, com variação na concentração de kefir de soja (K) e extrato
aquoso de soja (S): GS- 100% S; GK1- 75% S e 25% K; GK2- 50% S e 50% K; GK3- 25% S
e 75% K e GK- 100% K. Os produtos foram avaliados quinzenalmente, durante o período de
armazenamento de 90 dias a -15°C, sendo monitorados parâmetros microbiológicos
(viabilidade dos microrganismos potencialmente probióticos: Lactobacillus ssp., Streptococcus
spp., Bifidobacterium spp.; leveduras e segurança microbiológica: coliformes a 45°C;
Salmonella spp.) e sensoriais (teste de aceitação e intenção de compra). Todas as formulações
(GS, GK1, GK2, GK3 e GK) apresentaram ausência de Salmonella spp. e contagem de
coliformes a 45°C < 3 NMP/g durante todo o período de armazenamento, estando de acordo
com a RDC 12/2001. Nos tempos 0, 15 e 30 dias de armazenamento não foi detectada a
presença de leveduras nos gelados, enquanto a população de Bifidobacterium spp.,
Lactobacillus spp. e Streptococcus spp. foi superior a 10 log UFC/g. As formulações GK e GK2
apresentaram as maiores médias de aceitação para o atributo sabor (5,94 e 5,82,
respectivamente), não diferindo significativamente da GK3 e GS. Não foram verificadas
diferenças significativas entre as formulações para os atributos aparência (6,72 a 7,16), cor
(6,73 a 7,18) e aroma (5,51 a 5,98). Ademais, GK, GK3 e GK2 obtiveram as maiores
frequências de intenção de compra positiva, com 44,5%, 38,2% e 34,7 % dos consumidores,
afirmando que certamente ou provavelmente comprariam os produtos, respectivamente.
Conclui-se que foi possível obter um gelado comestível vegetal com adequada estabilidade
microbiológica durante o período de armazenamento e boa aceitação frente ao mercado
consumidor de produtos funcionais, mesmo com substituição total do EHS por kefir de soja.
Palavras-chave: sorvete, kefir, extrato aquoso de soja; probióticos, jaboticaba.
International Workshop on Advances of Probiotics and Prebiotics (IWAPP)
90
EFEITO DA DIETA COM NUCLEOTÍDEO NA RESPOSTA IMUNE INATA E
RESISTÊNCIA AO STREPTOCOCCUS AGALACTIAE EM TILÁPIA-DO-NILO